Identification and characteristics of Testudinis Carapax et Plustrum based on fingerprint profiles of mitochondrial DNA constructed by species-specific PCR and random amplified polymorphic DNA

• Published in: Mitochondrial DNA. Part B. Resources 2018-10, Vol.3 (2), p.1009-1012
• Main Authors: Li, Mingcheng, Wang, Miao, Zhou, Yuqing, Li, Zitong, Yuan, Guangxin, Wang, Xuesong, Xia, Wei, Chen, Jiayu
• Format: Article
• Language: English
• Subjects: cytochrome b; Mito Communication; mtDNA; PCR fingerprint profiles; Testudinis Carapax et Plustrum
• ISSN: 2380-2359; 2380-2359
• DOI: 10.1080/23802359.2018.1507631

Traditional use of Testudinis Carapax et Plustrum (TCP) as a medicine and health food has been widely reported. We compared two DNA fingerprint profiles of mitochondrial (mtDNA) from TCP based on species-specific PCR and random amplified polymorphic DNA (RAPD) to identify their authority. A series of sequences from cytochrome b (Cyt b) of Chinemys reevesii and their counterfeits were downloaded from the Genbank, and Premier 5.0 software was used to design a set of primers. A species-specific PCR and RAPD were undertaken to obtain the different DNA fingerprints respectively. The mtDNA was successfully extracted from all samples using the modified salting-out method. A relative molecular mass of 16.6 × 10 3 bp was observed, and mtDNA was measured between 1.83 ± 0.02. Fragments of 78 bp were amplified from all the TCP samples tested (except adulterant animals) using species-specific PCR method. The RAPD showed different electrocardiogram between genuine and counterfeit tortoise shell goods along with stripe number and location. The salting-out method (as modified) was used to extract high-quality mtDNA from TCP. The species-specific PCR and RAPD assay proposed in this study could be used for quality control of TCP with more specificity, sensitivity, and applicability.